The overall aim of this explorative proposal is to survey the microbiological and biochemical characteristics of a specific group of trypsin-like-enzyme (TLE) producing gliding organisms which are referred to in this proposal as "Rough Capnocytophaga" (RCC). These organisms have been found in the subgingival plaque of periodontitis patients involved in a longitudinal study in our laboratory. In addition, we would like to include other capnocytophage (smooth colony formers, SCC) which are capable of producing TLE and are prevalent in periodontitis and/or puberty gingivitis plaques of diabetic and non-diabetic children. At least 25 isolates of RCC from periodontic plaques, TLE producing reference strains from the American Type Culture Collection, and 30 to 40 isolates from puberty gingivitis plaques will be studied during the tenure of the proposal. Preliminary characterization of these organisms will be done using conventional microbiological tests including gas-liquid chromatographic analysis of glucose fermentation products. A battery of enzymes profile tests (aminopeptidases and glycosidases) for these organisms will be performed using rapid tests developed in our laboratory and the commercially available test systems, (AN-Ident, RapID ANA). Biochemical characterization of trypsin-like enzyme and its partial purification will be carried out by gel permeation and anion exchange columns using high-performance biocompatible fast protein liquid chromatography system (FPLC). The over-all information should eventually lead to a better understanding of the biochemical mechanisms and the role of these organisms in the pathogenesis of periodontal diseases. If so, a comprehensive research proposal will be planned in the future. The research facilities in laboratories of Oral Bacteriology and Biochemistry are excellent for the research work proposed in this application. Dr. Kauko Makinen, Director of the Biochemistry Laboratory will serve as a consultant in the enzyme characterization and purification study.